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首页> 美国卫生研究院文献>Applied and Environmental Microbiology >Culture-Dependent and Culture-Independent Characterization of Microbial Assemblages Associated with High-Temperature Petroleum Reservoirs
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Culture-Dependent and Culture-Independent Characterization of Microbial Assemblages Associated with High-Temperature Petroleum Reservoirs

机译：与高温石油储层相关的微生物组合的文化依赖性和文化依赖性

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Recent investigations of oil reservoirs in a variety of locales have indicated that these habitats may harbor active thermophilic prokaryotic assemblages. In this study, we used both molecular and culture-based methods to characterize prokaryotic consortia associated with high-temperature, sulfur-rich oil reservoirs in California. Enrichment cultures designed for anaerobic thermophiles, both autotrophic and heterotrophic, were successful at temperatures ranging from 60 to 90°C. Heterotrophic enrichments from all sites yielded sheathed rods (Thermotogales), pleomorphic rods resembling Thermoanaerobacter, and Thermococcus-like isolates. The predominant autotrophic microorganisms recovered from inorganic enrichments using H2, acetate, and CO2 as energy and carbon sources were methanogens, including isolates closely related to Methanobacterium, Methanococcus, and Methanoculleus species. Two 16S rRNA gene (rDNA) libraries were generated from total community DNA collected from production wellheads, using either archaeal or universal oligonucleotide primer sets. Sequence analysis of the universal library indicated that a large percentage of clones were highly similar to known bacterial and archaeal isolates recovered from similar habitats. Represented genera in rDNA clone libraries included Thermoanaerobacter, Thermococcus, Desulfothiovibrio, Aminobacterium, Acidaminococcus, Pseudomonas, Halomonas, Acinetobacter, Sphingomonas, Methylobacterium, and Desulfomicrobium. The archaeal library was dominated by methanogen-like rDNAs, with a lower percentage of clones belonging to the Thermococcales. Our results strongly support the hypothesis that sulfur-utilizing and methane-producing thermophilic microorganisms have a widespread distribution in oil reservoirs and the potential to actively participate in the biogeochemical transformation of carbon, hydrogen, and sulfur in situ.
机译：最近在不同地区对油藏的调查表明，这些生境可能具有活跃的嗜热原核生物组合。在这项研究中，我们使用了基于分子和基于文化的方法来表征与加利福尼亚州高温，富硫油藏相关的原核生物联盟。专为自养和异养厌氧嗜热菌设计的富集培养在60至90°C的温度下成功进行。来自所有部位的异养富集产生鞘杆（Thermotogales），类似于嗜热厌氧菌的多形杆和类似嗜热球菌的分离物。使用H2，乙酸盐和CO2作为能源和碳源从无机富集中回收的主要自养微生物是产甲烷菌，包括与甲烷杆菌，甲烷球菌和甲烷菌物种密切相关的分离株。使用古细菌或通用寡核苷酸引物组，从生产井口收集的总群落DNA中生成两个16S rRNA基因（rDNA）文库。通用文库的序列分析表明，很大一部分克隆与从相似栖息地回收的已知细菌和古细菌分离物高度相似。 rDNA克隆文库中代表的属包括嗜热厌氧杆菌，嗜热球菌，脱硫硫弧菌，氨基杆菌，酸氨基球菌，假单胞菌，嗜盐单胞菌，不动杆菌，鞘氨醇单胞菌，甲基杆菌和 Desulfomicrobium 。古细菌库中有大量类似甲烷菌的rDNA，而属于 Theocococcales 的克隆所占的百分比较低。我们的研究结果强有力地支持了以下假设：利用硫和产甲烷的嗜热微生物在油藏中分布广泛，并且有可能积极参与原位碳，氢和硫的生物地球化学转化。

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